Development and Clinical Validation of RT-LAMP-Based Lateral-Flow Devices and Electrochemical Sensor for Detecting Multigene Targets in SARS-CoV-2.
Apoorva SaxenaPawankumar RaiSrishti MehrotraSamiya BabySuman SinghVikas SrivastavaSmriti PriyaSandeep K SharmaPublished in: International journal of molecular sciences (2022)
Consistently emerging variants and the life-threatening consequences of SARS-CoV-2 have prompted worldwide concern about human health, necessitating rapid and accurate point-of-care diagnostics to limit the spread of COVID-19. Still, However, the availability of such diagnostics for COVID-19 remains a major rate-limiting factor in containing the outbreaks. Apart from the conventional reverse transcription polymerase chain reaction, loop-mediated isothermal amplification-based (LAMP) assays have emerged as rapid and efficient systems to detect COVID-19. The present study aims to develop RT-LAMP-based assay system for detecting multiple targets in N, ORF1ab, E, and S genes of the SARS-CoV-2 genome, where the end-products were quantified using spectrophotometry, paper-based lateral-flow devices, and electrochemical sensors. The spectrophotometric method shows a LOD of 10 agµL -1 for N, ORF1ab, E genes and 100 agµL -1 for S gene in SARS-CoV-2. The developed lateral-flow devices showed an LOD of 10 agµL -1 for all four gene targets in SARS-CoV-2. An electrochemical sensor developed for N-gene showed an LOD and E-strip sensitivity of log 1.79 ± 0.427 pgµL -1 and log 0.067 µA/pg µL -1 /mm 2 , respectively. The developed assay systems were validated with the clinical samples from COVID-19 outbreaks in 2020 and 2021. This multigene target approach can effectively detect emerging COVID-19 variants using combination of various analytical techniques at testing facilities and in point-of-care settings.
Keyphrases
- sars cov
- loop mediated isothermal amplification
- genome wide
- respiratory syndrome coronavirus
- copy number
- genome wide identification
- sensitive detection
- human health
- gold nanoparticles
- high throughput
- quantum dots
- risk assessment
- coronavirus disease
- molecularly imprinted
- gene expression
- highly efficient
- genome wide analysis
- label free
- visible light
- single cell